The long term goal of this research program is to understand how the phototransduction machinery is assembled, transported and maintained in outer segment (OS), as a prerequisite for preventing and/or repairing defects that arise when this process goes awry. We believe protein lipid modification plays a role in this process. We hypothesize that lipidation of proteins is a dynamic process and is essential in organizing and facilitating cross-talk between proteins in the phototransduction pathway at disc membranes. We will use phosophodiesterase-6 (PDE6) as a model protein to test this hypothesis. PDE6 is the crucial effectors enzyme needed for light signaling in rod and cone photoreceptor cells. Absence of PDE6 in ciliated OS in photoreceptor cells leads to vision loss accompanied by rapid degeneration. In humans, lack of PDE6 in OS leads to various blinding diseases such as retinitis pigmentosa, leber congenital amaurosis and achromatopsia. Despite our knowledge about the role of PDE6 as an effectors enzyme in phototransduction, how this crucial enzyme is assembled in inner segments, transported and then anchored in outer segment membranes is not known. The experiments proposed in this project are aimed at deciphering the mechanism behind the need for protein lipidation and further processing of PDE6 in survival and function of rods and cones. We plan to accomplish these goals by investigating the function, stability, assembly and transport of lipidated proteins including PDE6, in animal models that either expresses mutant forms of PDE6 or in models that lack specific enzymes that are responsible for these modifications. Finally, we will investigate the protein transport mechanisms in cone cells using these animal models. Our proposed studies are aligned with Retinal Diseases Program of the NEI to determine the pathophysiological mechanisms underlying mutations that cause retinal degenerative diseases. Our proposed studies lay a framework by which we can understand the basis behind various blinding diseases and design novel therapies to treat them.